U.S. patent application number 14/736339 was filed with the patent office on 2015-10-01 for methods for disinfecting or sterilizing articles.
The applicant listed for this patent is Joseph DUNN, PERFORMANCE PACKAGING OF NEVADA, LLC. Invention is credited to Joseph DUNN.
Application Number | 20150274397 14/736339 |
Document ID | / |
Family ID | 47830404 |
Filed Date | 2015-10-01 |
United States Patent
Application |
20150274397 |
Kind Code |
A1 |
DUNN; Joseph |
October 1, 2015 |
METHODS FOR DISINFECTING OR STERILIZING ARTICLES
Abstract
Disclosed herein are materials and methods for reducing,
preventing, or eliminating the biological contamination of surfaces
in spaces that are reasonably partitioned, defined or contained. A
reagent that includes peroxides or molecules that includes peroxide
bonds are contacted with at least one surface of an article. Some
of these methods include disinfection within an apparatus and may
include the step of volatilizing the peroxide bond containing
agent, through, for example, vapour pressure and vaporization or
evaporation effects, the addition of volatilization aids, or
passive or assisted diffusion at modest temperatures and relatively
long exposure times measured in some embodiments on the order of
hours, days, or months. In some embodiments a liquid that includes
a peroxide moiety is added (for example, during manufacture or
subsequently) to a package, container, or any other reasonably
partitioned or contained volume and allowed to migrate within the
confined space for a period of time sufficient to disinfect the
inner surfaces of the container and articles it contains.
Inventors: |
DUNN; Joseph; (Daytona
Beach, FL) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
DUNN; Joseph
PERFORMANCE PACKAGING OF NEVADA, LLC |
Daytona Beach
Las Vegas |
FL
NV |
US
US |
|
|
Family ID: |
47830404 |
Appl. No.: |
14/736339 |
Filed: |
June 11, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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13228105 |
Sep 8, 2011 |
9078435 |
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14736339 |
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Current U.S.
Class: |
206/368 ;
206/363; 206/440; 206/524.3 |
Current CPC
Class: |
A61B 2050/3008 20160201;
A61L 2/186 20130101; B65B 55/10 20130101; A01N 59/00 20130101; A61L
2202/23 20130101; B65D 81/18 20130101; A61B 50/30 20160201; B65D
25/14 20130101; B65D 25/04 20130101; A01N 59/00 20130101; B65B
55/02 20130101; A01N 25/18 20130101; A01N 2300/00 20130101; A01N
37/16 20130101 |
International
Class: |
B65D 81/18 20060101
B65D081/18; B65B 55/02 20060101 B65B055/02; A61B 19/02 20060101
A61B019/02; B65D 25/14 20060101 B65D025/14; B65D 25/04 20060101
B65D025/04 |
Claims
1-20. (canceled)
21. A container comprising: at least one wall defining an enclosed
space, the at least one wall having a surface in the enclosed
space, a chemical comprising a mixture of water, hydrogen peroxide
and formic acid inside the container, the at least one surface
having been exposed to the chemical in its vapor phase for at least
5 minutes at a temperature below 50.degree. C. whereby the at least
one surface is disenfected.
22. The container according to claim 21, further comprising an
article in the container, the article being selected from the group
of: a medical device, a dental or surgical instrument, an implant,
a foodstuff, labware, and gauze.
23. The container according to claim 21, wherein the container is
flexible.
24. The container according to claim 21, wherein the mixture
contains a hydrogen peroxide concentration equal to or greater than
about 2%.
25. The container according to claim 21, wherein the mixture
contains an acid concentration equal to or greater than about
2%.
26. The container according to claim 21, wherein the mixture
contains at least one additional supplement that increases or
elevates the vapor pressure of the aqueous mixture.
27. The container according to claim 21, wherein the concentration
and oxidizing capacity of the mixture diminishes over time and the
initial concentration or remaining residue from the mixture is too
low to pose an unacceptable threat to human or animal health.
28. The container according to claim 21, wherein the volume of
water, hydrogen peroxide and formic acid is less than 2.5 ml.
29. The container according to claim 21, wherein the enclosed space
includes partitioned spaces.
30. A container comprising: enclosed space for an article to be
disinfected, at least one surface within the enclosed space of the
container, a chemical which in the vapor phase is selected from the
group consisting of: peroxides, peracids, or moieties that include
at least one peroxide bond, a seal for sealing the container with
the chemical inside the container, wherein the at least one surface
within the container is disinfected by evolved gas from the mixture
after exposure for at least 1 hour at a temperature at or below
50.degree. C. and without exposing the mixture to microwave
radiation for forming a plasma.
31. The container according to claim 30, wherein the chemical
comprises an aqueous mixture of water, hydrogen peroxide and formic
acid.
32. The container according to claim 30, wherein the container is
rigid.
33. The container according to claim 30, wherein the container
includes an article disinfected by the chemical.
34. The container according to claim 33, wherein the article is
selected from the group consisting of: fitments, valves, a medical
device, a dental or surgical instrument, an implant, a foodstuff,
labware and gauze.
35. The container according to claim 30, wherein the concentration
and oxidizing capacity of the chemical diminishes over time and the
initial concentration or remaining residue from the chemical is too
low to pose an unacceptable threat to human or animal health.
36. The container according to claim 30, wherein the chemical
contains a hydrogen peroxide concentration equal to or greater than
about 2%.
37. The container according to claim 31, wherein the chemical
contains at least one additional supplement that increases or
elevates the vapor pressure of the aqueous mixture.
38. The container according to claim 30, wherein the container
includes partitioned spaces.
39. The container according to claim 38, wherein there is a first
article in a first of the partitioned spaces and a second article
in a second of the partitioned spaces.
40. A container providing contamination control comprising: a
container defining an enclosed space, an article to be disinfected
in the enclosed space, a chemical comprising a mixture of water,
hydrogen peroxide and formic acid inside the container with the
article, whereby the article is disinfected after having been
exposed to the chemical in its vapor phase for at least 5 minutes
at a temperature below 50.degree. C. and without exposing the
mixture to microwave radiation for forming a plasma.
Description
FIELD OF THE INVENTION
[0001] This invention relates generally to materials and methods
for sterilizing or disinfecting a variety of devices, articles and
materials; these methods can be practiced in situ and include
methods for eliminating or reducing the biological contamination of
the interior volume and surfaces of containers or structures.
BACKGROUND
[0002] Microorganisms, such as bacteria, fungi and the like are
virtually ubiquitous in the biosphere and while most are harmless,
many have the capacity to infect humans, plants and animals while
still others can cause spoilage of valuable foodstuffs and other
materials. Accordingly, there exists a pressing, ongoing need to
control, if not eliminate, unwanted microorganisms from a wide
variety of surfaces. One of the most common methods of disinfecting
or sterilizing surfaces is the use of heat, especially wet heat in
the form of high pressure steam. Unfortunately, these methods do
not work well with materials that are heat and/or water labile and
oftentimes it is difficult if not impossible to employ these
methods in situ.
[0003] For heat and/or moisture sensitive materials, alternatives
to dry heat and/or steam must be used to control the biological
contamination of these materials. Some of these alternatives
include, but are not limited to, irradiation, exposure to toxic
gases (for example, ethylene oxide, chlorine dioxide, or ozone),
ultraviolet radiation, or treatment with chemical bactericides,
fungicides and the like. Many of these methods are difficult to use
with various articles of manufacture so there remains a need for
new materials and methods for controlling microbial contamination
on surfaces and articles. Some aspects of the instant invention
address these needs.
SUMMARY
[0004] Some embodiments of the invention provide methods for
disinfecting and/or sterilizing the interior volume and surfaces of
sealed containers or structures using peroxides and molecules that
include peroxide bonds. Some embodiments provide methods for
disinfecting and/or sterilizing surfaces which leave behind small,
generally recognized as safe (GRAS) levels of chemical
residuals.
[0005] Some embodiments are methods performed using articles such
as packages, containers, bags, pouches, packaging, wrappings, tubs,
tins, bottles, buckets, and formed structures, or any other
article; preferably those articles having a reasonably partitioned
or defined space or volume within which a reagent that may include
a peroxide is inoculated, sprayed, or otherwise placed in contact
with at least one surface of the article. Some of these methods
include disinfection within an apparatus and may include the step
of volatilization of the peroxide bond containing agent, through,
for example, vapor pressure and evaporative effects, the use of
volatilization aids, or passive or assisted diffusion at modest
temperatures and providing relatively long exposure times, measured
in some embodiments on the order of hours, days, or months. In some
embodiments a liquid that includes a peroxide moiety is added (for
example, during manufacture or subsequently) to a package,
container, or any other reasonably partitioned volume and allowed
to migrate within the defined space for a period of time sufficient
to disinfect the inner surfaces of the container.
[0006] Some embodiments of the present invention also use as the
active agent performic acid (generated by the mixture of water,
formic acid, and hydrogen peroxide with or without volatilization
aids) for the control or elimination of microorganisms from
surfaces.
[0007] Some embodiments include methods for controlling
contamination within a partitioned or otherwise defined space,
comprising the steps of contacting surfaces within a partitioned or
otherwise defined space with a chemical in the vapor phase. In some
embodiments the chemical in the vapor phase is selected from the
group consisting of: peroxides, peracids, or moieties that include
at least one peroxide bond. In some embodiments the surfaces are
exposed to the chemical in its vapor phase for at least 5 minutes
at a temperature below 50 C. In some embodiments the partitioned or
otherwise defined space includes at least one article that is not
an integral part of the partitioned or defined space or volume.
[0008] In some embodiments the partitioned of otherwise defined
space is a container. In some embodiments the container is selected
from group consisting of: packaging, containers, bags, pouches,
cans, tins, buckets, tubs, bottles, ampoules, barrels, sleeves,
capsules, boxes, other formed packages or structures, and the like.
In some embodiments the containers are flexible. And in other
embodiments the containers are rigid. In some embodiments the
container may include at least one accessory or feature. In some
embodiments the accessory or feature is selected from the group
consisting of: fitments, valves, bungs, lids, vents, ports, spouts,
drains, covers and caps, seals, or membrane seals.
[0009] In some embodiments surfaces of or within a partitioned or
defined space or volume are incubated with a peroxide, peracid, or
a mixture of compounds, one or more of which includes a peroxide
bond for longer than about five minutes. In some embodiments the
temperature the incubation step occurs at a temperature equal to or
less than about 50 C. In some embodiments a peroxide concentration
equal to or greater than about 2% is added to or used in a
partitioned or defined space to form a peracid or a mixture of
compounds one or more of which includes a peroxide bond.
[0010] In some embodiments the peroxide mixture added to or used in
the partitioned or defined space includes formic acid (methanoic
acid) or performic acid (permethanoic acid) either of which may be
added to or present at a concentration greater than or equal to
about 2%. In some embodiments the peroxide mixture added to or used
in the partitioned or defined space includes peracetic acid
(ethanoic acid) or peracetic acid (perethanoic acid), or propionoic
acid (propanoic acid) or propionoic peracid (propanoic peracid), or
a mixture of these with or without the addition of formic acid, any
of or the combination of which is added or present at a
concentration greater than or equal to 2%. In some embodiments the
peroxide mixture added to or used in the partitioned or defined
space includes at the time of addition, or subsequently evolves to
form, a mixture that is generally recognized as safe for
consumption or other usage.
[0011] Still other embodiments include methods for reducing the
number of biological contaminants in a partitioned or otherwise
defined space or volume, comprising the steps of: mixing formic
acid, hydrogen peroxide and water with or without the addition of
other compounds; wherein the mixture produces performic acid in
situ; and contacting at least one surface with said performic acid
formed in situ wherein the surface is within a partitioned or
otherwise defined space or volume. In some of these embodiments the
space or volume is the interior of a container. In some embodiments
the container is selected from the group consisting of; packaging,
bags, pouches, cans, tins, boxes, buckets, tubs, bottles, ampoules,
barrels, sleeves, formed packages or structures or the like. In
some embodiments the partitioned or otherwise defined space or
volume includes at least one article and the article is present in
the space or volume at the same time as a microbiocidal
concentration of peracid or peroxide bond containing vapors. In
some embodiments the interior of the container includes at least
one article inside the container at the same time as a
microbiocidal concentration of peracid or peroxide bond containing
vapors.
[0012] Still other embodiments include systems for controlling
contamination within a partitioned or otherwise defined space or
volume comprising: a vapor that includes a microbiocidal
concentration of peracid or peroxide bonds within a partitioned or
otherwise defined space or volume, wherein the concentration of
vapors is such that at least about 5 minutes of contact between the
vapor and an interior surface of the space or volume is sufficient
to inactivate at least about 90 percent of the microbes within the
space or volume. In some embodiments the vapor is formed by a
mixture that includes peracid or peroxide formic acid (methanoic
acid) or performic acid (permethanoic acid) either of which is
added or present at a concentration greater than or equal to about
2%. In some embodiments the system is used to treat the interior of
a container which may or may not house an article that is not an
integral part of the container.
BRIEF DESCRIPTION OF THE FIGURES
[0013] FIG. 1A. Photograph of the inner surface of a fitment cap
showing the dried bacterial spore inoculum. The 10 microliter
inoculum dries to leave behind a circular, thick cream colored
spore mass.
[0014] FIG. 1B. Similar to 1A, a photograph of the inner surface of
a fitment cap showing the dried bacterial spore inoculum.
[0015] FIG. 2. The 10 microliter spore inoculum dried onto the
fitment shown in FIG. 1 viewed at about 25 times magnification
using a stercomicroscope.
[0016] FIG 3. Two 57.9 gallon drum bags. For testing, some bags
were incubated in a flat position (bottom bag), while other bags
were treated in a folded configuration (lower right hand corner)
after depositing a disinfectant mixture in the bottom of the bag in
order to demonstrate the ability of the disinfecting mixture to
migrate past the folds.
[0017] FIG. 4. Graph of the change in weight over time of
polyolefin bags containing water or a disinfecting mixture. The
weight loss is due to migration of water and/or components of the
disinfection mixture through the walls of the bag.
DESCRIPTION
[0018] For the purposes of promoting an understanding of the
principles of the novel technology, reference will now be made to
the preferred embodiments thereof, and specific language will be
used to describe the same. It will nevertheless be understood that
no limitation of the scope of the novel technology is thereby
intended; such alterations, modifications, and further applications
of the principles of the novel technology being contemplated as
would normally occur to one skilled in the art to which the novel
technology relates are within the scope of this disclosure and what
it claims.
[0019] Unless specifically stated or clearly implied the term
`about` as used herein refers to a range of values from less than
to greater than 10 percent of the stated value. For example, about
1 refers to the range of values from 0.9 to 1.1
[0020] Multiple methods and features of the present invention are
novel in relation to current practice. Though peroxide and peracids
are commonly used for disinfection and sterilization of containers
and packaging(s), they are most often employed so as to produce
rapid activity for on-line disinfection at relatively short
exposure times. The active agents are generally applied by
immersion, spraying, filling, or other methods designed to
distribute the active agents rapidly and throughout or over the
product to be disinfected or sterilized. The disinfection agents
are most often used at high concentration and volume, and/or with
the addition of a stimulant such as heat, ultraviolet light, a
plasma rich environment, or the like in order to activate the
oxidative activity of the peroxide-moiety and accelerate its decay
and attendant release of its oxidative capacity.
[0021] Contrary to currently used commercial methods the present
invention uses relatively small volumes of disinfecting reagents,
and relatively long times of exposure (for example, hours or days)
to produce disinfection or sterilization while yielding low levels
of residual compounds. The active agents employed in the present
invention are distributed throughout the treated product by
evaporation and other volatilization means followed by passive or
facilitated diffusion.
[0022] Some embodiments of the instant invention generate performic
acid in situ. In some embodiments performic acid mixtures are
produced by combining water, formic acid, and hydrogen peroxide
inside of the space that is to be disinfected. The utility of this
approach may be further extended by partitioning or confining the
space so as to allow the concentration of performic acid or vapors
from performic acid to accumulate in the given space. Given enough
time and the necessary amount of performic acid in a given
substantially confined or otherwise partitioned space, this reagent
can be used for the in situ sterilization of surfaces in a
partitioned space or volume.
[0023] Peroxide or peracid based reagents used for disinfecting or
sterilizing surfaces are usually used in combination with
accelerators of peroxide decomposition such as heat, ultraviolet
light, plasmas, or other activators which speed-up the formation of
hydroxyl radicals. In contrast, many of the embodiments of the
present invention can be practiced without the need for such
accelerants of peroxide decomposition. These methods are especially
useful for the disinfection and/or or sterilization of sealed
containers, such as packages, containers, bags, pouches, tanks,
tubs, bottles, buckets, sleeves or other formed structures, using a
small volume of hydrogen peroxide or another peracid agent, in
conjunction with long contact time. The peroxide bond rich vapor
migrates within the container and produces microbial disinfection
on the surfaces and the volume within the container through
oxidative effects acting over a relatively long time of exposure
(for example, hours or days rather than a few seconds or minutes)
at ambient or modestly elevated temperatures.
[0024] Some embodiments of the current invention are especially
well suited for the sterilization of bags for use in bag-in-box
(BIB) applications within the food and beverage industry.
Currently, bags for BIB applications are manufactured at one site
and then the bag/fitment combination is sterilized at a second site
using irradiation. Sterilization by irradiation in this process
usually requires that the bags be transported to a distant
irradiation facility, inserted into the irradiation processing
queue, and transported back to the originator for sale and
distribution following irradiation sterilization. This process is
expensive due to the required shipping and processing (the
irradiation process itself is relatively expensive in part because
of the high capital costs associated with the facility and
shielding of the radiation source), and time consuming (typically,
the minimum time required for the round trip is one to two weeks or
more).
[0025] In contrast, invention methods discussed herein can be used
virtually anywhere. In some embodiments of the invention a small
volume of a reagent that may include a peroxide or peracid mixture
is employed. The reagent may be sprayed, painted, or otherwise
dispensed into a bag, or a portion of the bag, or the neck of a
fitment that opens into the bag before, or during, the attachment
of the fitment to the bag. Volatilization of the active compounds
within the bag disinfects the inner volume and surfaces of the bag
including surfaces of the bag that are not in direct contact with
the liquid or solid form of the peroxide containing reagent. In
some embodiments, the active disinfection mixture may be added to a
fitment containing a membrane seal, and/or to a bag to which the
fitment is subsequently affixed. By controlling the volume of
disinfectant employed and/or continued disintegration of the active
chemistries, it is usually possible to generate harmless and
generally recognized as safe residuals and residual levels in the
bag at the time of filling. Accordingly, aspects of the present
invention provide relatively rapid and inexpensive methods
(relative to the transportation and processing times and costs
generally attendant to conventional radiation processing) for
disinfecting or sterilizing surfaces in confined spaces such as
bags. These methods are readily adaptable for BIB applications.
[0026] In still other embodiments these methods are used to
sterilize or disinfect medical products or devices packaged for use
at a later time, examples of such products, include, but are not
limited to scalpels or scalpel blades, luers, shunts, catheters,
sutures, and implants packaged in a wrap or container eventually
sealed to exclude re-contamination by microorganisms. The inclusion
of a small volume of disinfectant (according to some embodiments of
the present invention) before sealing of the package provides a
reagent for disinfecting or sterilizing the interior of the package
and the surfaces of any article(s) within the package. These
inventive methods enable the manufacturer to avoid costly radiation
or other energy intensive methods of sterilization or disinfection,
such as use of a retort.
Experiments and Exemplary Results
Experiment 1
[0027] About two or three droplets of 3% hydrogen peroxide were
droppered into a 4 ounce PET bottle before capping; the peroxide
was omitted from two bottles which were used as control samples.
Before sealing, the inside surface of the bottle cap (liner
removed) or its foam liner had been inoculated with a 10 .mu.L
droplet of Bacillus atrophaeus ATCC 9372 spores dried down onto the
cap or liner surface. Bacillus atrophaeus ATCC 9372 spores are
known to be of relatively high resistance to peroxide disinfection
and are a recommended (for example, the U.S. Pharmacopeia)
resistant organism for disinfection studies using chemicals (such
as peroxide or ethylene oxide) or dry heat.
[0028] After standing overnight (15 hours) at 35 C the bottles were
opened and the inoculated areas (of both untreated, control bottles
and bottles to which hydrogen peroxide had been added) were swabbed
to sample the level of recoverable viable spores. The results
obtained are shown in Table 1 below. The level of spores
recoverable from bottles to which 3 drops of 3% hydrogen peroxide
had been added was less (reductions >90% or >1 logarithm
lower) than that from control bottles to which no hydrogen peroxide
had been added. Thus, the hydrogen peroxide mixture used in this
experiment did reduce the level of recoverable viable spores and
show some disinfection capability, however, the level of efficacy
was small and insufficient to prove useful for the purposes of
sterilization.
TABLE-US-00001 TABLE 1 The recovery of Bacillus atrophaeus spores
from 4 oz. PET bottles incubated with (3 Drops, Overnight) or
without (None, Recovery Control) the addition of 3% hydrogen
peroxide. Site of CFU Log Log Inoculation Treatment Recovered
Recovery Reduction Liner None (Recovery 2300000 6.4 NA Control) Cap
None (Recovery 6600000 6.8 NA Control) Liner 3 Drops, 830000 6 0.4
Overnight Cap 3 Drops, 425000 5.6 1.2 Overnight Cap 3 Drops, 475000
5.7 1.1 Overnight
Experiment 2
[0029] A 10 microliter droplet of water containing Bacillus
atrophaeus spores was dried onto the inner surface of the fitment
from a Scholle Clean-Clic.RTM. 4L (BG 0004 PM 900/8CC21 WB)
bag-in-box bag. One milliliter of a performic acid mixture produced
by a combination of water, formic acid, and hydrogen peroxide was
placed in the bottom of the bag and the fitment then reinstalled;
replicate inoculated fitments were placed on bags without the
addition of performic solution in order to serve as control
samples. The bags were stored at ambient conditions (approximately
65-75.degree. F.) for one week, the inoculation site on each
fitment was then swabbed, the swabs shaken in 3 mL of a saline
solution to harvest the recovered spores, and the harvest medium or
its serial ten-fold dilutions spread plated on tryptic soy agar
medium and incubated for 48 hours at 35 C. The number of viable
Bacillus atrophaeus spores recovered from untreated (Recovery
Control) or performic treated bags is shown in Table 2.
TABLE-US-00002 TABLE 2 The number of colony forming units (CFU)
recovered from fitments of bags incubated without (Recovery
Control) or with the addition of a performic mixture. Log Sample
CFU Recovered Log CFU Recovered Reduction Recovery Control 63800
4.8 NA Recovery Control 4300 4.6 NA Treated 0 <0 >4.7 Treated
0 <0 >4.7
[0030] As illustrated by the data summarized in Table 2, no viable
spores were recovered from the performic treated bag sample
fitments, while 46,000 CFU or more of spores were recovered from
the fitments on untreated, control bags.
Experiment 3
[0031] As in Experiment 2, a 10 microliter droplet of water
containing Bacillus atrophaeus spores was dried on the inner
surface of the fitment from a Scholle Clean-Clic.RTM. 4 L (BG 0004
PM 900/8CC21 WB) bag-in-box bag. The inoculum level used was
significantly greater than was used in Experiment 2. Referring now
to FIGS. 1A, 1B, and 2. FIGS. 1A and 1B, which show inoculated
fitments on which a 10 microliter water droplet containing greater
than 10,000,000 (10 million) colony forming units of Bacillus
atrophacus spores was dried. The spore inoculation site on each
fitment was clearly visible after drying (FIGS. 1A and 1B); clearly
the spot contains a very high concentration of spores. The area of
the spot is about 0.4 cm.sup.2 and it contains greater than
10,000,000 spores (a spore concentration greater than 25 million
spores per square centimeter). This is a relatively high level of
inoculation and the spore mass at the inoculation site is clearly
visible. Referring to FIG. 2. The 10 microliter water droplet dried
onto the fitment shown in FIG. 1, viewed at about 25 times
magnification using a stereomicroscope. Note the thick outer ring
of spores formed as the droplet dried; a portion of this thick
outer ring has dislodged from the upper part of the dried droplet
during handling of the fitment before viewing. This outer ring
contains a thick biomass of spores deposited one on top of the
other many spores deep.
[0032] About one milliliter of a performic acid mixture produced by
a combination of water, formic acid, and hydrogen peroxide was
placed in the bottom of the bag and the fitment then reinstalled;
replicate inoculated fitments were placed on bags without the
addition of performic solution in order to serve as control
samples. The bags were stored at ambient conditions (approximately
65-75.degree. F.) for 20, 44, and 68 hours and the inoculation site
on each fitment was then swabbed, recovered, and counted as in
Experiment 2. The number of viable Bacillus atrophaeus spores
recovered from untreated (Recovery Control) or performic treated
bags is shown in Table 3 below. Some surviving spores were
recovered from one of the treated samples after 20 hours of
treatment (but not the replicate sample), but no viable spores were
recovered on any of the treated samples thereafter. About 7.5
logarithm cycles of viable spores (or more than 10 million CFU)
were recovered from the untreated, control samples.
TABLE-US-00003 TABLE 3 The number of colony forming units (CFU)
recovered from fitments of bags incubated without (Recovery
Control) or with the addition of 1 mL of a performic mixture.
Treated samples were assayed after 20, 44, and 68 hours; the
control samples were assayed at 20 and 68 hours. Log CFU Sample CFU
Recovered Recovered Log Reduction Recovery Control 34300000 7.5 NA
Recovery Control 31400000 7.5 NA 20 Hrs 225 2.4 5.1 20 Hrs 0 <0
>7.5 44 Hrs 0 <0 >7.5 44 Hrs 0 <0 >7.5 68 Hrs 0
<0 >7.5 68 Hrs 0 <0 >7.5
Experiment 4
[0033] As in Experiments 2 and 3, a 10 microliter droplet of water
containing Bacillus atrophaeus spores was dried onto the inner
surface of the fitment from a Scholle Clean-Clic.RTM. 4 L (BG 0004
PM 900/8CC21 WB) bag-in-box bag. The inoculum level used was high
as in Experiment 3. For treatment, 300 microliters or 100
microliters of a performic acid mixture produced by a combination
of water, formic acid, and hydrogen peroxide was placed in the
bottom of a bag and the fitment then reinstalled; a replicate
inoculated fitment was placed on a bag without the addition of
performic solution in order to serve as a control sample. The bags
were stored at ambient conditions (approximately 65-75.degree. F.)
for 50 hours and the inoculation site on each fitment was then
swabbed, recovered, and counted as in Experiments 2 and 3. The
number of viable Bacillus atrophaeus spores recovered from
untreated (Recovery Control) or performic treated bags is shown in
Table 4 below. No surviving spores were recovered from either of
the treated samples. About 7.7 logarithm cycles of viable spores
(or more than 10 million CFU) were recovered from the untreated,
control sample.
[0034] After the 50 hour incubation at ambient conditions, sample
bags treated with 100 microliters of the performic solution were
filled with 3 liters of water, shaken, and then sampled and assayed
for peroxide residuals using CHEMetrics, Inc. (4295 Catlett Rd.,
Calverton, Va. 20138) Chemets K-5510 assay procedures. The peroxide
residuals were 1 part per million (ppm, mg/L) in the water sample.
A peroxide residuals reading was also taken on a bag treated with
100 microliters of the performic solution and assayed after 9 days
of incubation at ambient conditions. Again the bag was filled with
3 liters of water, shaken, and then sampled and assayed as before.
The peroxide residuals were now between 0.6 and 0.8 ppm in the
water sample.
TABLE-US-00004 TABLE 4 The number of colony forming units (CFU)
recovered from fitments of bags incubated without (Recovery
Control) or with the addition of either 300 microliters or 100
microliters of a performic mixture. Samples were assayed after 50
hours at room temperature. Log Sample CFU Recovered Log CFU
Recovered Reduction Recovery Control 54100000 7.7 NA 300 microL, 50
Hrs 0 <0 >7.7 100 microL, 50 Hrs 0 <0 >7.7
Experiment 5
[0035] As in Experiments 2, 3, and 4, a 10 microliter droplet of
water containing Bacillus atrophaeus spores was dried on the inner
surface of the fitment from a Scholle Clean-Clic.RTM. 4L (BG 0004
PM 900/8CC21 WB) bag-in-box bag. The inoculum level used was a
one-to-ten dilution of that used in Experiments 3 and 4. For
treatment, 25 microliters of a performic acid mixture produced by a
combination of water, formic acid, and hydrogen peroxide was placed
in the bottom of a bag and the fitment then reinstalled; a
replicate inoculated fitment was placed on a bag without the
addition of performic solution in order to serve as a control
sample. The bags were stored at ambient conditions (approximately
65-75.degree. F.) for 46 and 122 hours and the inoculation site on
each fitment was then swabbed, recovered, and counted as previously
described. The number of viable Bacillus atrophaeus spores
recovered from untreated (Recovery Control) or performic treated
bags is shown in Table 5 below. Surviving spore recovery from
either of the treated samples was significantly less than that from
the untreated, control samples.
[0036] After the 33 hours at ambient conditions incubation, a
sample bag treated with 25 microliters of the performic solution
was filled with 3 liters of water, shaken, and then sampled and
assayed for peroxide residuals using the Chemets K-5510 assay
procedures. The oxidative peroxide residuals were 0.3 parts per
million (ppm) in the water sample. A second oxidative peroxide
residuals reading was taken after 122 hours of incubation at
ambient conditions. Again the bag was filled with 3 liters of
water, shaken, and then sampled and assayed as before. The
oxidative peroxide residuals level was now 0.2 ppm in the water
sample. The regulatory limit for oxidative peroxide residual levels
under these conditions is 0.5 ppm; clearly the disinfection process
yielded peroxide oxidative levels well below the regulatory
limit.
TABLE-US-00005 TABLE 5 The spore recovery from fitments of bags
incubated without (Recovery Control) or with the addition of either
25 microliters of a performic mixture. Samples were assayed after
46 and 122 hours at room temperature. Log Sample CFU Recovered Log
CFU Recovered Reduction Recovery Control 2,600,000 6.4 NA Recovery
Control 2,650,000 6.4 NA 25 microL, 46 Hrs 2280 3.4 3.0 25 microL,
122 Hrs 8 0.9 5.5
Experiment 6
[0037] The effect of concentration of the added disinfection
mixture was investigated by comparing 500 microliter of the
disinfection mixture used in the previous demonstrations (call this
mixture a 1.times. concentration of the disinfecting agent) and 200
microliters of a mixture containing 3.8 times the level of peroxide
as the 1.times. concentration mixture.
[0038] As in many of the previous exhibits, a 10 microliter droplet
of water containing Bacillus atrophaeus spores was dried on the
inner surface of the fitment from a Scholle Clean-Clic.RTM. 4L (BG
0004 PM 900/8CC21 WB) bag-in-box bag. For treatment, each of the
two volumes and concentration mixture combinations of water, formic
acid, and hydrogen peroxide were placed in the bottom of a bag and
the fitment then reinstalled; a replicate inoculated fitment was
placed on a bag without the addition of performic solution in order
to serve as a control sample.
[0039] The results obtained from this experiment are summarized in
Table 6. These data illustrate that by elevating the concentration
of the disinfection mixture added to the package it is possible to
accelerate the migration/permeation of the disinfection mixture
within the bag.
TABLE-US-00006 TABLE 6 The effect of the concentration of the
disinfection mixture on the time required to produce disinfection
and sterilization within a treated bag. Untreated, Control 7.3
Logarithm Cycles of Colony Forming Units Recovered 1X Concentration
500 .mu.L for 6.6 Logarithm Cycles of Colony Forming 17 Hours Units
Recovered 3.8X Concentration 200 .mu.L 3 Colony Forming Units
Recovered for 17 Hours
Experiment 7
[0040] Demonstration tests were performed to document the ability
of this method to effectively disinfect and sterilize even larger
packages than the 4 liter bags employed previously. Fifty seven and
nine-tenths (57.9) gallon bags were used for these tests (Scholle
200301 BG 0220 APHM 800/800X). For these tests, a 10 microliter
droplet of water containing Bacillus atrophaeus spores was dried on
the inner surface of the fitment and also onto the inner surface of
the bag immediately beneath the fitment.
[0041] Five milliliters or 2.5 mL of the disinfection mixture
(1.times. concentration) was placed in the bag as low and as far
below the fitment as possible and the bag held upright and shaken
slightly in order to cause the disinfection mixture to run to the
bottom of the bag. One bag to which 5 mL of disinfection mixture
was added and one bag to which 2.5 mL was added were incubated at
ambient temperatures in an unfolded flat position, and one bag to
which 5 mL of disinfection mixture was added was folded before
incubation (see FIG. 3).
[0042] The bags were incubated for thirteen days and the results
obtained for the number of viable Bacillus atrophaeus spores
recovered from an untreated (Control) bag or the bags to which the
disinfection mixture had been added are summarized in Table 7
below.
TABLE-US-00007 TABLE 7 The numbers of viable colony forming units
recovered from 57.9 gallon drum bags. Innoculation Site Sample
Fitment Bag Control 6.73 6.72 Log CFU Log CFJ 5 mL of 1X 0 0
Concentration CFU CFU 2.5 mL of 1X 0 0 Concentration CFU CFU Folded
Bag 0 0 CFU CFU
[0043] Referring now to FIG. 3. For testing two 57.9 gallon drum
bags were incubated in a flat position or after careful folding
(attempting to retain the added disinfection mixture in the bottom
of the bag). After swab recovery of the Bacillus atrophaeus spores
from the two inoculation sites on the disinfection mixture treated
bags, the bags were filled with 57.5 gallons (483 pounds) of water,
mixed, and then sampled and assayed for peroxide residuals using
the Chemets K-5510 assay procedures. The oxidative peroxide
residual determinations obtained are shown in Table 8.
TABLE-US-00008 TABLE 8 The oxidative residual levels obtained from
57.9 gallon drum bags after 13 days. 5 mL of 1X 0.3 mg/L
Concentration 2.5 mL of 1X 0.1 mg/L Concentration Folded Bag 0.4
mg/L (5 mL of 1X)
Experiment 8
[0044] This exhibit is an extension of the methods and results
shown in Experiment 7 using the same 57.9 gallon drum bags used in
Experiment 7 (Scholle 200301 BG 0220 APHM 800/800X). The volumes
and disinfection mixture concentrations employed are shown in Table
9. The tested volumes and concentrations of the disinfection
mixture were able to inactivate all the inoculated Bacillus
atrophaeus spores after incubation for about four (4) days at
ambient conditions. Note that in these trials the disinfection
mixture was deposited at the foot end of the bag opposite the
fitment, the bag folded so as to retain the disinfection mixture at
that end of the bag during the folding process, and the bag was
then incubated in this folded position; the disinfection mixture
was nevertheless able to still migrate across the folding pattern
to inactivate spores inoculated on the fitment. Greater than 6
(6.4) logarithm cycles of viable spore colony forming units were
recovered from the untreated, control bag.
[0045] Referring now to Table 9. In both trials the disinfection
mixture was deposited at the foot end of the bag opposite the
fitment, the bag folded, and the bag treated in a folded position.
Also, in these particular tests the oxidative potential of the
residual contents of the bag after filling with 5.3 gallons of
water were below the regulatory maximum (regulatory limit is 0.5
mg/L) as the level of recovered residuals in these tests were 0.1
and 0.2 mg/L. These results show that it is possible to sterilize
these large bags (57.9 gallons) during room temperature storage in
as short a time as four days and with negligible and generally
recognized as safe levels of residuals.
TABLE-US-00009 TABLE 9 Recovery of viable spores and oxidative
residuals from 57.9 gallon drum bags after about 4 days of ambient
incubation. 95.5 Hrs 4 mL 0 CFU Recovered Residuals Log Red
Concentration 4.5 X 0 CFU Recovered 0.1 mg/L >6.4 95 Hrs 6 mL 0
CFU Recovered Residuals Log Red 96 Hrs Control 6.53 Log CFU
Recovered 96 Hrs Control 6.32 Log CFU Recovered
Experiment 9
[0046] The previous exhibits employed bags with relatively high
barrier properties, i.e., the disinfectants and steriliants added
were confined to the bag, the materials and construction of the
bags being relatively inhibitory to the migration of moisture or
gasses through the walls of the bag.
[0047] In this experiment, a polyolefin bag (Scholle 2.6 gallon
200119 BG 0010 PP 900/1400 WB) with significantly lower barrier
properties (a bag relatively more permeable to moisture vapor or
gasses) was employed to demonstrate the efficacy of the method even
under such conditions and with such materials wherein the treated
container is not strictly hermetic nor the treated volume strictly
confined.
[0048] During initial trials, adding up to even 1 milliliter of the
disinfection mixture at the 1.times. concentration yielded no
significant Bacillus atrophaeus spore inactivation even after 55
hours of ambient incubation. Subsequent trials using 1.5 mL and 2.5
mL of the 1.times. concentration of the disinfection mixture
yielded 2 logarithm cycles or less inactivation after 141 hours
(>5.8 days) of ambient incubation using a 6 logarithm cycle
inoculation level of Bacillus atrophaeus spores.
[0049] After these poor initial results, a weight loss study was
performed. Referring now to FIG. 4. The graph in FIG. 4 illustrates
the loss of weight when 1.5 mL of water, 1.5 mL of the disinfection
mixture at the 1.times. concentration, or 5 mL of the 1.times.
concentration disinfection mixture were sealed into the bag and
incubated at ambient conditions inside a sealed chamber containing
Drierite (W.A. Hammond Drierite, Ltd, Xenia, Ohio). An
approximately 0.013 to 0.015 grams per hour loss in weight from the
bag was observed. The results obtained clearly demonstrate that
during incubation, compounds in the bag are migrating through the
packaging material and being lost to the environment surrounding
the bag.
[0050] The migration through the bag (effusion) necessitates an
adjustment of the composition and amount of disinfection mixture
needed to be added to the bag in order to achieve sterilization.
Follow on experiments examined the effects of using different
concentrations and volumes of the disinfection mixture. As in
previous experiments, a 10 microliter droplet of water containing
Bacillus atrophaeus spores was dried on the inner surface of the
fitment from each of a duplicate series of bags. In each of the
test bags, the volume of disinfection mixture at the concentration
listed in Table 10 was deposited near the center of the bag, the
fitment replaced on each bag, and the bag incubated for 23 or 25
hours at ambient temperatures (see Table 10), at which time the
inoculation site on each fitment was swab recovered and viable
spores enumerated.
[0051] The results summarized in Table 10 illustrate that in these
2.6 gallon polyolefin bags a range of disinfection mixture
concentrations and added volumes were able to inactivate all or
nearly all of the Bacillus atrophaeus spores within about a day of
incubation at ambient temperatures and conditions. These results
clearly demonstrate that an appropriate volume and concentration of
the disinfection mixture can be used to disinfect and sterilize the
internal volume of polyolefin bags.
TABLE-US-00010 TABLE 10 The disinfection and sterilization effects
of using various volume and concentration combinations to treat 2.6
gallon polyolefin bags. (Log Red is a convenient abbreviation for
Logarithm Cycle Reduction in Spore Viability). 25 Hrs 200 .mu.L 0
CFU Recovered Residuals Log Red Concentration 2.7 X 0.15 mg/L
>7.31 23 Hrs 200 .mu.L 0 CFU Recovered Residuals Log Red
Concentration 4.5 X 0.3 mg/L >7.31 25 Hrs 150 .mu.L 6.64 Logs
Recovered Residuals Log Red Concentration 1.3 X 0.05 mg/L 0.66 23
Hrs 150 .mu.L 0 CFU Recovered Residuals Log Red Concentration 2.2 X
0.0 mg/L >7.31
Experiment 10
[0052] This exhibit is an extension of the methods and results
shown in Experiment 9 to a second, larger a polyolefin bag (Scholle
200233 BG 0020 PP 900/1400). The volumes and disinfection mixture
concentrations employed are shown in Table 11. Again it was seen
that the selected volume and concentration of the disinfection
mixture was able to inactivate all the inoculated Bacillus
atrophaeus spores after one day of ambient incubation. Note that in
one trial the disinfection mixture was deposited at the foot end of
the bag opposite the fitment, the bag folded so as to retain the
disinfection mixture at that end of the bag during the folding
process, and then the bag incubated in this folded position; the
disinfection mixture was nevertheless still able to migrate (across
the folding pattern) to inactivate spores inoculated on the
fitment. Greater than 7 (7.14) logarithm cycles of viable spore
colony forming units were recovered from the untreated, control
bag.
[0053] Referring now to Table 11. In these particular tests the
oxidative potential of the residual contents of the bag after
filling with 5.3 gallons of water were about twice the allowable
regulatory maximum (regulatory limit is 0.5 mg/L; recovered
residual levels in these tests were 1.0 mg/L).
TABLE-US-00011 TABLE 11 The disinfection and sterilization effects
of using various volume and concentration combinations to treat 5.3
gallon polyolefin bags. 24 Hours 400 mL 0 CFU Recovered Residuals
Log Red Concentration 4.5X 0.1 mg/L >7.14 24 Hours Repeat 400 mL
0 CFU Recovered Residuals Log Red Concentration 4.5X 0.1 mg/L
>7.14 24 Hours Folded 400 mL 0 CFU Recovered Residuals Log Red
Concentration 4.5X Folded 0.1 mg/L >7.14
[0054] While the novel technology has been illustrated and
described in detail in the figures and foregoing description, the
same is to be considered as illustrative and not restrictive in
character, it being understood that only the preferred embodiments
have been shown and described, and that all changes and
modifications that come within the spirit of the novel technology
are desired to be protected. As well, while the novel technology
was illustrated using specific examples, theoretical arguments,
accounts, and illustrations, these illustrations and the
accompanying discussion should by no means be interpreted as
limiting the technology. All patents, patent applications, and
references to texts, scientific treatises, publications, and the
like referenced in this application are incorporated herein by
reference in their entirety.
* * * * *